The ongoing revolution in
aging research was manifested by the Second International Conference
"Genetics of Aging and Longevity" (Moscow, April 22-25, 2012). It was
organized by the Science for Life Extension Foundation in collaboration with the
Gerontological Society of the Russian Academy of Sciences and the International Association of Gerontology and Geriatrics (European Region). More
than 250 gerontologists and and other researchers in the area of aging from 27
countries (Austria, Armenia, Belarus, Brazil, Germany, Greece, Denmark, Israel,
India, Iran, Italy, Kazakhstan, Canada, Latvia, Lithuania, Nigeria,
Netherlands, Poland, Portugal, Puerto Rico, Russia, United Kingdom, USA,
Ukraine, France, South Korea, Japan) gathered to discuss current problems of
genetics, life expectancy, and aging mechanisms. Among these scientists were
invited speakers Vladimir Anisimov, Vladislav Baranov, Andrzej Bartke, Nir
Barzilai, Mikhail Blagosklonny, Holly Brown-Borg, Judith Campisi, Vadim
Fraifeld, Claudio Franceschi, David Gems, Vadim Gladyshev, Vera Gorbunova,
Andrei Gudkov, Valter Longo, Brian Kennedy, Brian Kraemer, Kyung-Jin Min,
Richard Morimoto, Alexey Moskalev, Thomas Perls, Suresh Rattan, Blanka Rogina,
Andrei Seluanov, Robert Shmookler Reis, Yousin Suh, Nektarios Tavernarakis, Jan
Vijg, Anatoli Yashin, and Piotr Zimniak. The Conference goal was to identify
the most promising areas of genetics, life expectancy, and aging, including:
the search for longevity genes; the search for pharmacological agents that slow
aging; the identification of biological age markers; and the identification of
mechanisms by which the environment influences the aging rate.

Judith Campisi (Buck Institute for Research on Aging,
USA) presented "Cancer and Aging: Rival Demons?" Cellular senescence is a
potent tumor suppressive mechanism that halts the proliferation of cells at
risk for neoplastic transformation. Senescent cells accumulate with
age and at sites of age-associated pathology in many tissues. There
is mounting evidence that senescent cells are not mere bystanders; rather, they
contribute to aging phenotypes and age-related disease and thus may be an
example of evolutionary antagonistic
pleiotropy. One strong candidate is the senescence-associated
secretory phenotype (SASP). The SASP entails the transcriptional
upregulation and secretion of numerous cytokines, growth factors and proteases
that can have profound paracrine effects. The SASP can disrupt the
structure and function of normal tissues. It can also, ironically,
provoke malignant phenotypes in premalignant cells. Thus, the SASP may be
the mechanism by which senescent cells drive both the degenerative and
hyperplastic pathologies of aging. What controls the
SASP? They identified three signaling pathways that are important
positive regulators of the SASP. These are the DNA damage response
pathway, the p38MAPK-NFkB pathway, and, recently, the mTOR
pathway. mTOR signaling is of particular interest because it ties
the SASP to an evolutionary conserved pathway that regulates longevity in
diverse species, ranging from yeast to mice. Why did complex
organisms evolve a senescence response and a SASP? There is little
doubt that the senescence growth arrest evolved to suppress the development of
cancer. The SASP, by contrast, may have evolved to promote tissue
repair. Results from a new mouse model developed by the group
strongly indicate that this is indeed the case for skin
wounds. Thus, the senescence response subserves three processes:
tumor suppression, tissue repair, and age-related pathology, including,
ironically, cancer.

Brian Kennedy (Buck Insitute for Research on Aging,
USA) presented "The TOR pathway and aging - insights from genetic screens".
Evidence from invertebrate model organisms has indicated that many pathways
modulating aging are conserved. Among the most conserved is the TOR pathway,
which acts as a cellular monitor of nutrient levels in the environment.
Reduced TOR signaling promotes stress resistance and enhanced longevity in
yeast, worms, flies and mice - the most studied aging model organisms. Current
work is aimed at understanding the pathways downstream of TOR that contribute
to aging. Rapamycin, a specific mTOR inhibitor confers robust extension of
lifespan and healthspan in mice, offering promise that it will be possible to
develop therapeutic agents aimed at extending healthspan. Currently,
derivative of rapamycin are being tested in the Kennedy lab to enhance
protection against age-related diseases while simultaneously reducing side
effects. Increasingly, it seems possible that it will be possible to intervene
in the aging process and the TOR pathway is among the most promising targets.

Mikhail V. Blagosklonny (Roswell Park Cancer Institute, Buffalo,
USA) presented a report "mTOR from cellular senescence to organismal aging".
The nutrient-sensing TOR (Target of Rapamycin) pathway is involved in cellular
and organismal aging. Pharmacological interventions that inhibit this pathway
increase life span in yeast, nematode, Drosophila and rodents. In a number of
experiments, rapamycin suppressed and slowed down cellular senescence of human
cells. The mTOR inhibitor rapamycin prevents age-related weight gain, decreases
rate of aging, increases life span and decreases carcinogenesis in mice.

David Gems (University College London, U.K.)
presented "Does hyperfunction cause aging in C. elegans?" Many recent studies
have failed to support the view that oxidative damage causes aging in C.
elegans. One possibility is that the stochastic damage-somatic maintenance
paradigm of aging is incorrect. An alternative proposed mechanism involves
quasi-programmed hyperfunction (manifested e.g. as hypertrophy). There are many
signs of hypertrophy in aging C. elegans, e.g. yolk over-production and
cuticular thickening. Quasi-programmed apoptosis simultaneously causes gonadal
disintegration (atrophy) and oocyte gigantism (hypertrophy) in aging C. elegans
hermaphrodites. Hermaphrodite specificity of physiological germline apoptosis
accounts for sex differences in pathology of aging in the gonad. The
hyperfunction theory provides a full and simple account for a major age-related
pathology in C. elegans. Hyperfunction is a viable alternative to molecular
damage as a central mechanism of aging in C. elegans.

Vladimir Anisimov (N.N. Petrov Research Institute of
Oncology, Russia) in his presentation "Do we really have a medicine against
aging?" showed results of experiments on
effects of antidiabetic biguanides and rapamycin on biomarkers of aging, life
span, spontaneous and chemically-induced carcinogenesis in outbred, inbred and
transgenic HER-2/neu mice and in rats. The mTOR inhibitor rapamycin prevents
age-related weight gain, decreases rate of aging, increases life span and
decreases carcinogenesis in transgenic HER-2/neu cancer-prone mice. Rapamycin
dramatically delayed tumors onset, decreased a number of tumors per animal and
tumor size. Lifelong administration of rapamycin extends lifespan in female
129/Sv mice characterized by normal mean lifespan of 2 years. Importantly,
rapamycin was administrated inter-mittently (every other 2 weeks) starting from
the age of 2 months. Rapamycin inhibited age-related weight gain, decreased
aging rate, increased lifespan (especially in the last survivors) and delayed
spontaneous cancer. 22.9% of rapamycin-treated mice survived the age of death
of the last mouse in control group. Treatment of female outbred SHR mice with
metformin started at the age of 3 months increased mean life span by 14% and
maximum life span by 1 month. Same treatment started at the age of 9 months
insignificantly increased mean life span by 6%, whereas treatment started at
the age of 15 months failed to increase life span. When started at the age of 3
and 9 months, metformin delayed time of the first tumor detection by 22% and
25%, correspondingly. Thus, in female SHR mice, metformin slowed down aging and
somewhat postponed appearance of tumors when started at the young and middle
but not at the old age. Yet, metformin improved reproductive function when
started at any age. The chronic treatment of inbred 129/Sv mice with metformin
slightly modified the food consumption but failed to influence the dynamics of
body weight, decreased by 13.4% the mean life span of male mice and sligtly
increased the mean life span of female mice. The treatment with metformin failed
influence spontaneous tumor incidence in male 129/Sv mice and decreased by 3.5
times the incidence of malignant neoplasms in female mice. Metformin increased
mean life span by 8% and mammary adenocarcinoma (MAC) latency by 13.2%
(p<0.05) in transgenic HER2/neu mice. The treatment metformin inhibited the
growth of transplantable HER2 mammary carcinoma in FVB/N male mice by 46% at
the 45th day after transplantation (p<0.001). We also have shown that
phenformin or metformin inhibits 1,2-dimethylhydrazine-induced colon
carcinogenesis in rats, X-rays, N-nitrosomethylurea- or
7,12-dimethylbenz(a)anthracene-benzo(a)pyrene-induced mammary carcinomas in
rats, cervicovaginal, skin and soft tissue tumorigenesis in mice. These results
suggest that both metformin and rapamycin may be useful in prevention of cancer
and extension of lifespan when used in rational and appropriate ages, doses and
schedules.

Andrei Gudkov's
(Roswell Park Cancer
Institute, Buffalo, USA) presentation was
focused on a recent discovery made in his lab of a new role of p53 as a
regulator of epigenetic silencing of non-coding part of mammalian genome and
deregulation of this major p53 function in cancer and aging.

Andrzej Bartke (Southern Illinois University School of
Medicine, USA) presented "Longevity benefits of endocrine defects". Mutations
which interfere with the production of, or with actions of, growth hormone (GH)
increase life expectancy in mice by as much as 60%. An important
implication of this finding is that the average and maximal lifespan are not
fixed characteristics of a species but can be altered by mutation of a single
gene. These long-lived mutant mice remain healthy and vigorous much
longer than their genetically normal siblings and are partially protected from
cancer. Results available to date indicate that reduced GH signaling
slows aging and extends longevity by multiple interacting
mechanisms. These mechanisms include increased stress resistance,
improved insulin signaling, reduced mTOR activity, alterations of metabolism
and mitochondrial function, a shift of secretory profile of adipose tissue from
pro- to anti-inflammatory, improved maintenance of stem cell populations and
organ-specific changes in expression of
numerous genes. Interestingly, it was recently reported that
genetically GH resistant humans never get diabetes and never die of
cancer. Moreover, people from exceptionally long-lived families have
improved insulin sensitivity. Improved insulin signaling and other
characteristics associated with extended longevity of GH-resistant and
GH-deficient mice can be safely induced in humans by physical activity and a
modest reduction of food intake.

Nir Barzilai (Einstein College of Medicine, USA)
presented "Lipid profiles and genotypes associated with exceptional longevity
in humans". We assessed phenotype and genotype of over 540 subjects with
exceptional longevity and their families. The families of the longest living
subjects are very distinct by their lipid profile (High HDL levels and large
lipoprotein particle sizes). However no major role of lifestyle factor was
noted in these subjects as the rates of overweight, smoking, physical activity
where generally worse over that reported in their cohort. On the other hand,
there are several genotypes that have been significant in such subjects,
including those in the GH/IGF pathway, lipid metabolism, thyroid metabolism,
and FOXO3A that have been confirmed in other groups. These findings indicate
that the genome of subjects with exceptional longevity is significantly
different than those who die at a younger age, and that this genome has been
protecting them against some of the worst environmental factors.

Alexey Moskalev (Institute of Biology at Komi Science
Center, RAS, Russia) presented "Drosophila melanogaster life extension by
overexpression of the Growth Arrest and DNA Damage-45 (GADD45) gene". The GADD45 protein family plays an important
role in stress signaling and participates in the integration of cellular
response to environmental and physiological factors. GADD45 proteins
are involved in cell cycle control, DNA repair and demethylation, apoptosis,
cell survival and senescence, and inflammatory response by complicated
protein-protein interactions. Because of their pleiotropic action, a decreased
indelibility of Gadd45 members may have far-reaching consequences including
genome instability, accumulation of DNA damage, and disorders in cellular
homeostasis - all of which may eventually contribute to the aging process and
age-related disorders (promotion of tumorigenesis, immune disorders, insulin
resistance and reduced responsiveness to stress). In Drosophila
melanogaster a single D-GADD45 ortholog has been described. Our
data show that overexpression of the D-GADD45 gene in the nervous system
leads to a significantly increase of Drosophila lifespan without a decrease in
fecundity and locomotor activity. The lifespan extension effect is more
pronounced in males than in females, which agrees with the sex-dependent
expression of this gene. The longevity of D. melanogaster with
D-GADD45 overexpression is apparently due to more efficient recognition
and repair of DNA damage, as the DNA comet assay showed that the spontaneous
DNA damage in the larva neuroblasts is reduced with statistical significance. Although
further wide-scale research is warranted, it is becoming increasingly clear
that Gadd45s are highly relevant to aging, age-related diseases (ARDs) and to
the control of life span, suggesting them as potential therapeutic targets in
ARDs and pro-longevity interventions.

Suresh Rattan (Aarhus University, Denmark) presented
"Ageing interventions: anti-ageing or prohealthspan?" If ageing is understood
as an emergent phenotype due to the failure of homeodynamics and not due to the
action of any harmful and death-causing mechanisms, it transforms our approach
towards ageing interventions from "anti-ageing" to "healthy ageing". Ageing
occurs in spite of the presence of complex pathways of maintenance, repair and
defence, and there is no "enemy within". This viewpoint makes modulation of
ageing different from the treatment of one or more age-related diseases.
Another important implication of understanding ageing as the inefficiency and
imperfections of homeodynamics is that the prospect of developing anti-ageing
magic bullets must be abandoned. This also means abandoning enemy-oriented
rhetoric, such as the "war against ageing", "defeating ageing", and "conquering
ageing" etc. Instead, interventions in ageing require a "friend-oriented"
approach and the use of a positive language such as maintaining health,
achieving healthy ageing, successful ageing, and preserving the homeodynamics.
A promising strategy to slow down ageing and prevent or delay the onset of
age-related diseases is that of mild stress-induced hormesis and hormetins.
Physical, nutritional and mental hormetins which initiate stress responses and
strengthen the homeodynamics are potentially effective ageing modulators. As a
biomedical issue, the biological process of ageing underlies all major
diseases, and although the optimal treatment of every disease, irrespective of
age, is a social and moral necessity, preventing the onset of age-related
diseases by intervening in the basic process of ageing is the best approach for
achieving healthy ageing and extending the health-span.

Robert Shmookler Reis (University of Arkansas for Medical
Sciences & VA Medical Center, USA), studying a set of isogenic longevity
mutants in C. elegans, discovered numerous genes for which transcript
abundances provide powerful predictors of longevity. In addition, lipid
characteristics (Peroxidation Index, Fatty Acid Chain Length, %MUFAs, %PUFAs)
are also valuable biomarkers of adult lifespan, and are consistent with
elongase & desaturase transcript levels. Proteome features, especially
KINASE profiles, reflect signaling pathway activities. These biomarkers often
translate to mice and/or humans. In another study of Quantitative Trait Loci
(QTLs) that naturally modify longevity as different strains evolve, Dr.
Shmookler Reis identified 27 highly significant loci that impact lifespan,
Darwinian fitness, and/or resistance to a variety of stresses. Of these QTLs,
13 had marked effects on longevity, and provided evidence that (1.)
antagonistic pleiotropy (counter?vailing effects of a locus on fitness vs.
lifespan) is common, but not universal (seen for about a third of loci); (2.)
only a few dozen genes are responsible for the evolutionary divergence of
longevity among natural isolates; and (3.) gene-gene interactions are quite
common, and may chiefly affect fitness or longevity; 3 interactions (of 9
observed) provided clear and significant evidence of antagonistic pleiotropy
acting at the level of gene interaction. One of the longevity loci has
recently been pursued to the point of identifying the gene responsible, which
turned out to be a gene involved in meiotic chromosome pairing. The allele
that stabilized the genome more during meiosis was associated with shorter
adult lifespan and lower stress resistance, again implying a trade-off between
reproductive benefits vs. post-reproductive survival.

Jan Vijg (Einstein College of Medicine, USA) presented "Aging of
the genome". Genomes of somatic cells are
highly dynamic and accumulate a considerable number of alterations over the
life span (genetic damages, mutations, epimutations). High-throughput
sequencing is an effective way to comprehensively characterize the aging genome
of individual somatic cells. Random genetic alteration may cause aging through
an increase in transcriptional noise. Epimutations in DNA methylation profiles
are frequent and well above the background of non-conversion events. The
epimutation frequency tend to increase with age. Epimutation frequency may vary
across the genome.

Claudio Franceschi (University of Bologna, Italy) presented "High throughput technologies
(OMICS) as a new tool to identify biomarkers of longevity and healthy aging".
The phenotype of centenarians is unexpectedly complex being a mixture of
adaptive robustness and accumulating frailty. Centenarians do have genetic risk
alleles for major age-related diseases despite their remarkable capability to
postpone / avoid them. They identified 4 chromosomal regions linked to
longevity (14q11.2, 17q12-q22, 19p13.3-p13.11, 19q13.11-q13.32). This is the
first study that detects linkage with longevity at the APOE gene locus.
Longevity is associated with mtDNA haplogroup J and higher levels of C150T
mutation in the Control Region (D-loop). A significant decrease in global DNA
methylation levels was observed with age (higher methylation percentage in
young controls compared to all other groups). Age-related loss of DNA
methylation was less pronounced in centenarians offspring, than in offspring of
non long-lived parents. N-glycans profiling appears to be one of the more
robust biomarker of healthy aging.

Richard Morimoto (Northwestern Center for Genetic
Medicine, USA) presented "The stress of misfolded proteins in biology, aging
and disease". Proteins are the major constituents of our cells; they
comprise the molecular machines that replicate the genome, transcribe RNA, and
translate other proteins. To achieve this uncoding of biological information,
each constituent protein must fold into a specific three-dimensional shape.
This process of protein folding is essential, and yet error-prone synthesis,
genetic polymorphisms, and physiological and environmental stress leads to
misfolding and the accumulation of damaged proteins. The balance between
function and dysfunction is achieved by cell stress response pathways (the heat
shock response and the unfolded protein response) and the proteostasis network
that ensures synthesis, folding, translocation, assembly, and clearance of the
proteome. During stress and aging, however, this balance is disrupted leading
to collateral damage and the amplification of protein damage with enhanced risk
for age-associated diseases including, cancer, immunological disease, metabolic
disease, and neurodegeneration. Therefore, key to healthspan and the prevention
of these diseases is an understanding of the underlying biology of protein
quality control, how damaged proteins are detected, and mechanisms of the HSR
and other stress responses to enhance folding and clearance of damaged
molecules.

Brian Kraemer (University of Washington & VA
Medical Center, USA) presented "Suppressing aging related proteotoxicity:
targeting tau pathology in C. elegans". Age dependent accumulation of
protein aggregates is one of the neuropathologies associated with brain
aging. Indeed deposition of aggregated detergent insoluble proteins
in neurons is a common feature in most aging dependent neurodegenerative
disorders. Abnormal deposits of human tau protein are the most
common pathological hallmarks of aging related dementia disorders including
Alzheimer's disease and frontotemporal dementia. Using a
transgenic C. elegans model for human tau pathology, we have
been exploring the genetic requirements for tau neurotoxicity. In
transgenic C. elegans expressing human tau in all neurons, we
observe several hallmarks of human tauopathies including altered behavior,
reduced lifespan, accumulation of detergent insoluble phosphorylated tau
protein and neurodegeneration. To identify genes required for
tau neurotoxicity, we have utilized forward genetic screens for mutations that
suppress tau neurotoxicity and associated phenotypes. We have isolated several
different mutations ameliorating the toxic effects of tau. While
these mutations alter a variety of processes in neurons including: autophagy,
nucleo-cytoplasmic transport, RNA processing, and microtubule function, the
exact mechanisms of neuroprotection remain unclear. Nonetheless,
all suppressor mutations isolated to date reduce tau aggregation and ameliorate
neurodegenerative changes. We will present new work on the
conservation of tau suppressor genes from worms to humans and the role these
homologous human genes play in the formation of pathological
tau. Likewise the potential for novel neuroprotective and life-span
extending strategies will be explored.

Vera Gorbunova and
Andrei Seluanov (University of Rochester, USA) presented "Aging and genome
maintenance". Genomic instability is a hallmark of aging tissues. Genomic
instability may arise due to inefficient or aberrant function of DNA
double-strand break (DSB) repair. DSBs are repaired by homologous recombination
(HR) and nonhomologous DNA end joining (NHEJ). HR is a precise pathway, while
NHEJ frequently leads to deletions or insertions at the repair site. Our
studies showed that NHEJ declines approximately 3-fold in replicatively
senescent cells. We also observed reduction in NHEJ efficiency in aging mice.
However, the strongest changes were observed with HR, which declined 38-fold in
pre-senescent cells. Expression of multiple proteins involved in HR diminished
with cellular senescence. Supplementation of these proteins either individually
or in combination did not rescue senescence related decline of the HR.
Remarkably, overexpression of SIRT6 in pre-senescent cells strongly stimulated
HR repair. These studies suggest that activation of SIRT6 may reduce
age-related genomic instability. Andrei Seluanov and Vera Gorbunova (University
of Rochester, USA) also presented "Longevity and anticancer mechanisms in
long-lived mole rats". They identified a novel mechanism of
cancer-resistance in the naked mole rat termed early contact inhibition
(ECI). Contact inhibition is a key anticancer mechanism that arrests
cell division when cells reach high density. Naked mole rat cells
are hypersensitive to contact inhibition and arrest proliferation at low cell
density. They found that ECI requires the activity of p53 and Rb pathways
and is associated with induction of p16INK4a. Recently
they identified the extracellular matrix component that that triggers ECI.
Application of this molecule in humans opens new avenues for cancer prevention
and life extension.

Sean Curran (University of Southern California, USA)
presented "A conserved starvation response mediated by non-canonical SKN-1/
Nrf2 signaling". SKN-1/Nrf2 plays multiple essential roles in development and
maintaining cellular homeostasis. We demonstrate a novel cellular mechanism
utilized by SKN-1 to execute a specific and appropriate transcriptional
response to changes in available nutrients leading to metabolic
adaptation. We isolated the first gain-of-function(gf) alleles of
skn-1, which disrupt the association of SKN-1 with the mitochondrial outer
membrane protein PGAM-5. In the presence of plentiful food, the
skn-1(gf) mutants perceive a state of starvation. The aberrant
monitoring of cellular nutritional status leads to an altered survival response
to changes in food availability. As such, skn-1(gf) mutants
transcriptionally activate genes associated with metabolism, adaptation to
starvation, aging, and survival. The triggered starvation response is conserved
in mice with constitutively activated Nrf2. The dysregulated
metabolism in these cells may contribute to the lethal phenotype and the
tumorgenicity associated with activating Nrf2 mutations in mammalian somatic
cells. Taken together, our findings provide a mechanism for an
evolutionarily conserved metabolic axis of SKN-1/Nrf2 activation that adds
further dimensions to the complexity of this pathway.

Dr. Holly Brown-Borg (University of North Dakota, USA)
presented "Growth hormone and aging: Does DNA methylation play a role?" Growth
hormone mutant mice are diminutive in size, exhibit enhanced antioxidative
capacity and extended longevity when compared to their growth hormone
sufficient counterparts. Many physiologic mechanisms appear to be altered in
these mice that may contribute to these differences including aspects of
glutathione and methionine metabolism. The atypical methionine metabolism in
the Ames dwarf mice leads to differential expression of DNA methylation enzymes
(Dnmt) and differential methylation. Gene expression of the Dnmt enzymes (Dnmt1,
Dnmt3a) is greater in Ames dwarf mice in comparison to age-matched wild type
mice (p<0.05). However, protein levels of Dnmt1 are lower while levels of
Dnmt3a protein tend to be higher in dwarf compared to wild type mice. Dnmt
enzyme activity levels also differ by genotype and age and may be responsible
for the differences in methylation observed in an earlier array experiment.
Global DNA hypomethylation was measured using repetitive DNA elements and found
to differ by genotype and age. In addition to the enhanced ability to counter
oxidative stress, these epigenetic changes may contribute to altered gene
expression and the overall extension of health span and lifespan enjoyed by
these mice.

Bertrand Friguet (Université Pierre et Marie Curie,
France) presented "Pathways affected by oxidative proteome modifications in
cellular senescence and oxidative stress". Accumulation of oxidized proteins is
a hallmark of cellular aging. Oxidized proteins, proteins modified by lipid
peroxidation and glycoxidation adducts have been previously shown to accumulate
in senescent human fibroblasts WI-38 (Ahmed et al., 2010, Aging Cell,
9:252-272). By using two-dimensional electrophoresis and immunoblotting coupled
with mass spectrometry, proteins targeted by these modifications have been
identified and found to include proteins mainly involved in protein
maintenance, energy metabolism and cytosketon. A similar proteomic approach has
also been used to identify oxidized protein targets in WI-38 fibroblasts
undergoing stress-induced premature senescence and only a restricted set of
proteins was found targeted by carbonylation. Changes in the proteome of human
myoblasts during replicative senescence and upon oxidative stress have been
recently analyzed. The carbonylated proteins identified either upon oxidative
stress (Baraibar et al., 2011, Free Rad. Biol. Med., 51:1522-32) or during
replicative senescence are involved in key cellular functions, such as
carbohydrate metabolism, protein maintenance, cellular motility and homeostasis.
Interestingly, almost half of the proteins identified as increasingly
carbonylated were the same during replicative senescence or upon oxidative
stress. Taken together, our results indicate that proteins involved in key
cellular pathways are affected upon oxidative stress and cellular senescence,
the impairment of which may be implicated in cellular dysfunction. Moreover,
this study underscores the importance of performing proteomic analyses
addressing different aspects, such as expression level and carbonylation, to
have a broader view of changes affecting the cellular proteome.

Blanka Rogina (University of Connecticut Health
Center, USA) presented "Mechanisms of Indy life extension in D.
melanogaster". Indy encodes the fly homologue of a mammalian transporter
of di and tricarboxylate components of the Krebs cycle intermediates.
Reduced expression of fly Indy or two of worm Indy homologs extend longevity.
In flies, INDY is predominantly expressed in places where intermediary
metabolism takes place, such as gut, fat body and oenocytes. It has been
hypothesized that Indy mutation mimics calorie restriction and extend longevity
by related mechanism. This hypothesis is supported by the similarities in
physiology of calorie restricted flies with Indy mutant flies on high calorie
diet, such as lower weight, egg production, levels of triglyceride, decreased
starvation resistance and increased spontaneous physical activity. In addition,
Indy mutant flies have similar changes in mitochondrial biogenesis as calorie
restricted animals. New findings also suggest that Indy mutation preserve
homeostasis in other tissues that contribute extended health and longevity in
Indy mutant flies.

Nektarios Tavernarakis (FORTH, Medical School, University of
Crete, Greece) presented "Mitochondrial energy metabolism and protein
homeostasis in aging". Regulation of protein synthesis is critical for
cell growth and maintenance. Ageing in many organisms, including humans, is
accompanied by marked alterations in both general and specific protein
synthesis. Whether these alterations are simply a corollary of the ageing process
or have a causative role in senescent decline remains unclear. A battery of
protein factors facilitates the tight control of mRNA translation initiation.
The eukaryotic initiation factor 4E (eIF4E), which binds the 7-methyl guanosine
cap at the 5' end of all nuclear mRNAs, is a key regulator of protein
synthesis. In addition, marked alterations in cellular energy metabolism are a
universal hallmark of the ageing process. The biogenesis and function of
mitochondria, the energy-generating organelles in eukaryotic cells, are primary
longevity determinants. Genetic or pharmacological manipulations of
mitochondrial activity, profoundly affect the lifespan of diverse organisms.
However, the molecular mechanisms regulating mitochondrial energy metabolism during
ageing are poorly understood. Prohibitins are ubiquitous, evolutionarily
conserved proteins, which form a ring-like, high molecular weight complex at
the inner membrane of mitochondria. Prohibitin function has been implicated in
carcinogenesis and replicative senescence. The nematode Caenorhabditis elegans
offers a versatile platform in which to investigate the potential link between
protein quality control, mitochondrial energy metabolism and ageing. We have
found that the mitochondrial prohibitin complex promotes longevity by
moderating fat metabolism and energy production in C. elegans.
Prohibitin deficiency shortens the lifespan of otherwise wild type animals.
Remarkably, knockdown of prohibitin promotes longevity under dietary
restriction, in diapause mutants and in animals under metabolic stress.
Depletion of prohibitin influences ATP levels, animal fat content and
mitochondrial proliferation in a genetic-background- and age-specific manner.
Together, these findings reveal a novel mechanism regulating mitochondrial
biogenesis and function, with opposing effects on energy metabolism, fat
utilization and ageing. Prohibitin may serve a similar key role in the
modulation of energy metabolism during ageing in mammals. Moreover, we find
that loss of a specific eIF4E isoform, IFE-2 that functions in somatic tissues,
reduces protein synthesis and extends the lifespan of C. elegans.
Knockdown of the phosphatidyl inositol kinase TOR that controls protein
synthesis in response to nutrient cues further increases the longevity of ife-2
mutants. In addition, reduction of protein synthesis increases ATP availability
and stress resistance Thus, signaling via eIF4E may influence ageing by
moderating energy demands and augmenting stress resistance mechanisms, though
regulation of protein synthesis in the soma.

Thomas Perls (Boston University, USA) presented
"Increasing genetic influence upon exceptional longevity with older and older
ages". Working with a wide range of disciplines including statisticians,
geneticists and computer scientists, has led the production of a landmark
article in which a genetic model consisting of 281 genetic markers predicts
with 85% accuracy whom in their sample of controls and centenarians is age 105+
years (published this January in PLoS ONE). The accuracy of the model is lower,
about 60% for nonagenarians and centenarians at age 100, which supports the
hypothesis that the genetic component of survival to older and older age beyond
100 gets progressively stringer.

Olga Mustafina (Institute of Biochemistry and Genetics,
RAS, Russia) presented "Gene polymorphism and human longevity: Association
studies deliver". In men FOXO1A gene polymorphism (rs4943794)
is possibly associated with ageing, and FOXO3A gene
polymorphism (rs3800231) may be important for achievement of longevity.

Elena Pasyukova (Institute of Molecular Genetics, RAS,
Russia) presented "Drosophila melanogaster lifespan is associated with genes
controlling asymmetric neuroblast division".One of the organ systems that play
a major role in maintaining homeostasis and regulating aging and longevity is
the nervous system. Earlier, we
demonstrated that several genes participating in regulation of the nervous system development are involved in lifespan
control. An insertion of the P{GT1} vector 300 bp downstream of the
structural part of escargot (esg) that encodes an RNA polymerase
II transcription factor and insertions of P-based vectors in the
structural part of aPRC that encodes atypical protein kinase C were
associated with increase in male and female lifespan. Both esg and aPKC
are involved in regulation of a crucial step in the nervous system development,
asymmetric neuroblast division (AND). We suggested that other genes interacting
with esg and aPKC during AND could be important for lifespan
control. inscrutable (insc) is a gene essential for AND. We
demonstrated that an insertion of the P{EPgy2} vector in the structural
part of insc prolonged female lifespan. Analysis of lifespan of
heterozygous esg and insc double mutants allowed us to predict
that these genes interact in the course of lifespan determination. Decrease of esg
transcription was shown to be associated with lifespan increase in mutant esg
and insc flies. aPKC is directly phosphorylated by GSK3β (glycogen
synthase kinase 3) encoded by shaggy (sgg), which further
provides AND. We demonstrated that several insertions of P-based vectors
in the structural part of sgg were associated with alterations of male
and female lifespan. Altogether, our recent findings indicate that genes
affecting AND during early steps of development could influence longevity of Drosophila
adults.

Piotr Zimniak (University Arkansas for Medical Sciences
& VA Medical Center, USA) presented "The lipid peroxidation product
4-hydroxynonenal as a modulator of fat accumulation and aging". Several
4-HNE-metabolizing enzymes and chemical 4-HNE scavengers have a longevity
assurance function in C. elegans. 4-HNE may accelerate aging either by its
toxicity as an electrophile or by modulating metabolism (ectopic fat
accumulation?) or signaling pathways such as insulin-like signaling. The
effects of 4-HNE on longevity and on fat deposition could be linked by a
metabolic syndrome-like state. 4-HNE-linked mechanisms are amenable to
pharmacological treatment. The oxidative stress theory of aging is not quite
dead yet, but is in need of chemical refinement.

Isabelle Petropoulos (Université Pierre et Marie Curie,
France) presented "Role of oxidized protein repair in protection against
oxidative damage: a proteomic approach". The accumulation of oxidatively
modified protein is a hallmark of aging. This accumulation results, at least in
part, from the increase of reactive oxygen species coming from both cellular
metabolism and external factors including environment, but the efficacy of
protein maintenance systems is also involved. Most organisms, from bacteria to
humans, have developed a specific reductase system, the Methionine Sulfoxide
Reductase (Msr), which allows the repair of oxidized methionines in proteins.
The Msr system, composed of the two stereo specific enzymes: MsrA and MsrB,
plays a major role in the maintenance of protein homeostasis during aging and
has also been involved in cellular defences against oxidative stress, by
scavenging ROS. To analyse more precisely the relationships between the Msr
system, oxidative stress and oxidative protein damage, MsrA and MsrB2, a
mitochondrial member of the MsrB family, have been stably overexpressed in
cellular models. Msr overexpressing cells are more resistant to H2O2 cytotoxity
by delaying apoptosis and protecting against necrosis (Cabreiro et al., 2008, J
Biol Chem, 283: 16673-16681). Moreover, it was demonstrated that the mechanisms
by which MsrB2 protects against oxidative stress include: maintenance of a
lower level of intracellular ROS, prevention of oxidized protein accumulation
and protection of the proteasome against oxidative stress induced inactivation.
Finally, stable human embryonic kidney cell lines (HEK) where MsrA, MsrB1 or
MsrB2 gene expressions are silenced by using RNAi technology, were generated.
These knocked down clones have been used in 2D-DIGE experiments (2D
fluorescence difference gel electrophoresis). Most of the proteins found to be
differentially expressed in Msr depleted cells were identified by mass
spectrometry as proteins related to redox homeostasis. Altogether, it is
suggested that the Msr proteins, in addition to be oxidized protein repair
enzymes, may play an important role in protein homeostasis and cellular
anti-oxidant defenses.

Mikhail Shaposhnikov (Insttute of Biology, Komi Science
Center, RAS, Russia) in his presentation "Drosophila life extension by
inhibitors of PI3K, TOR kinase and NFkB" have reported that recent progress in
our understanding of genetic mechanisms of aging and longevity provides an
opportunity to select some enzymes as targets for pharmacological intervention
into aging speed. Phosphoinositide 3-kinase (PI3K) and TOR-kinase cascades are
affected in some long-lived mutants of different animals, such as nematodes and
mice. NF-κB transcription factor is one of the major regulators of gene
expression associated with mammalian aging. The purpose of this study was to
investigate the anti-aging potential of the inhibitors of enzymes that have
known association with aging and longevity. Experimental Drosophila imagoes
were exposed to 5 μM of LY294002, 0.5 μM of wortmannin,
0.5 μM of rapamycin, and 120 μM of pyrrolidine
dithiocarbamate (PDTC) during their lifetimes. We used LY294002 and wortmannin
as specific PI3K inhibitors, rapamycin as the TOR-kinase inhibitor,
and PDTC as the NF-κB inhibitor. LY294002 treatment of Drosophila imago
has led to an increase of the median (by 14%) and maximum (by 16-22%) lifespan
(p<0.001) in females and males, respective-ly. Wortmannin
treatment has induced an increase of the median (by 5%) and maximum (by 39%)
lifespan in males (p<0.001), but the lifespan differences in females
were statistically insignificant (p>0.05). Rapamycin treatment has
induced increases of median (by 5-6%) lifespan (p<0.01) in males and
females, respectively and increase of maximum lifespan (by 33%) in females (p<0.01).
Treatment with PDTC has increased the median (by 11-13%) and the maximum (by
11-14 %) lifespan in females and males, respectively. Thus, we have shown that
the specific pharmacological inhibition of PI3K (by LY294002 and wortmannin),
TOR-kinase (by rapamycin), and NF-κB transcription factor (by PDTC) prolonged the Drosophila
melanogaster lifespan.

Vadim Fraifeld (Ben-Gurion University of the Negev,
Israel) presented "Linking cellular senescence and age-related diseases via
miRNA regulation of protein interaction networks". The comprehensive data
mining revealed over 250 genes tightly associated with cellular senescence
(CS). Using systems biology tools, it was found that CS is closely
interconnected with aging, longevity and age-related diseases (ARDs), either by
sharing common genes and regulators (miRNAs) or by protein-protein interactions
and eventually by common signaling pathways. The patterns of evolutionary
conservation of CS and cancer genes showed a high degree of similarity,
suggesting the co-evolution of these two phenomena. Moreover, cancer genes and
microRNAs seem to stand at the crossroad between CS and ARDs. The analysis
also provides the basis for new predictions: the genes common to both cancer
and other ARD(s) are highly likely candidates to be involved in CS and vice
versa. Multiple links between CS, aging, longevity and ARDs suggest a
common molecular basis for all these conditions. Modulating CS may represent a
potential pro-longevity and anti-ARDs strategy.

Anatoly Yashin (Duke University, USA) presented
"Mechanisms of aging and longevity: Lessons from genetic analyses of
longitudinal data". The associations of candidate genes with lifespan detected
in a number of studies were corroborated in some but not confirmed in other
studies. Attempts to replicate associations of genetic factors with lifespan
detected in earlier studies using the Genome Wide Association Studies (GWAS)
were not very successful: none or a very small fraction of the previously
detected associations have been confirmed. The genetic variants detected in GWA
studies of complex traits, including lifespan, explain only a small portion of
the narrow sense heritability that was calculated for such traits in the
pre-genomic era. This situation requires an explanation and indicates the need
for developing new approaches to the analysis of genetics of aging and
lifespan. We discuss one such approach, which combines GWAS of human lifespan
with analyses of longitudinal data on changes in health status and
physiological state. The approach addresses the issue of "missing heritability"
and allows for studying joint influence of many small-effect genetic variants
on lifespan and other durations (e.g., free of major diseases and disability
lifespan) by constructing polygenic score indices and investigating their
influence on such traits. It also allows for investigating differences in age
trajectories of physiological indices, as well as health histories for
individuals with different genetic background. We also show why conventionally
used strategy to confirm GWAS research findings often fails to do
so.

Vadim Gladyshev (Brigham and Women's Hospital, Harvard
University Medical School, USA) presented "Exceptional longevity of the naked
mole rat: insights from genome sequencing and transcriptomic
analysis". The naked mole rat (NMR, Heterocephalus glaber) is a strictly
subterranean, extraordinarily long-lived eusocial mammal. Although the size of
a mouse, its maximum lifespan exceeds 30 years and makes this animal the
longest living rodent. NMRs show negligible senescence, no age-related increase
in mortality, and high fecundity until death. In addition to delayed aging,
NMRs are resistant to both spontaneous cancer and experimentally induced
tumorigenesis. NMRs pose a challenge to the theories that link aging, cancer
and redox homeostasis. Although characterized by significant oxidative stress,
the NMR proteome does not show age-related susceptibility to oxidative damage
nor increased ubiquitination. NMRs naturally reside in large colonies with a
single breeding female, the "queen," who suppresses the sexual maturity of her
subordinates. NMRs also live in full darkness, at low oxygen and high carbon
dioxide concentrations, and are unable to sustain thermogenesis nor feel
certain types of pain. We report sequencing and analysis of the NMR genome,
which revealed unique genome features and molecular adaptations consistent with
cancer resistance, poikilothermy, hairlessness, altered visual function,
circadian rhythms and taste sensing, and insensitivity to low oxygen. This
information provides insights into NMR's exceptional longevity and capabilities
to live in hostile conditions, in the dark and at low oxygen. The extreme
traits of NMR, together with the reported genome and transcriptome information,
offer unprecedented opportunities for understanding aging and advancing many
other areas of biological and biomedical research.

Yelena Budovskaya (University of Amsterdam, Holland)
presented "Molecular aging driven by Wnt signaling in C. elegans." We
are using a system biology approach to reveal the molecular basis for aging in
nematodes C. elegans by characterizing gene expression
differences between young and old animals and then determining at a molecular
level how these changes contribute to aging. We used DNA microarrays to profile
gene expression changes associated with aging. This analysis revealed that gene
expression differences between young and old animals are under control of a
relatively simple gene regulatory network that involves the elt-3, elt-5,
and elt-6 GATA transcription factors. Expression of elt-5 and elt-6 increases
in old age, leading to decreased expression of elt-3, thus causing
changes in the expression of the many downstream target genes. We found no
evidence that it is caused by cellular damage or environmental stresses.
Rather, we found that elt-3 expression in the adult is controlled
by increased expression of the repressors elt-5 and elt-6,
which also guide elt-3 expression during development. These
results suggest that age-regulation of elt-3 is caused by
age-related drift of an intrinsic developmental program that becomes imbalanced
in old age. This pathway, which is one of
the first and clearest examples of developmental drift, may in part be
responsible for some of the
age-related changes that occur as the worm grows old. A key unanswered question
is what causes elt-3/elt-5/elt-6 transcriptional drift during
aging? In C. elegans, Wnt/Wingless signaling pathways
activate elt-5 and elt-6 expression during
development. Here we demonstrated that Wnt signaling is responsible for the
increased expression of elt-5 and elt-6 GATA
transcription factors during aging. Mutations in the components of the
"Wnt/b-catenin asymmetry pathway", such us mom-2/Wnt,
the β-catenin, wrm-1, or pop-1/TCF decreases
expression levels of the elt-5 and elt-6 GATA
transcription factors throughout life, leading to increased elt-3 GATA
expression. We demonstrated that inactivation of the "Wnt/b-catenin asymmetry
pathway" starting at the firtst day of adulthood, extends lifespan of otherwise
wild type animals by ~40 - 50%. These results indicate that changes in Wnt
signaling - a regulator of the elt-3/elt-5/elt-6 GATA
transcriptional circuit during normal development - plays an important role in
the age-related regulation of the elt-3/elt-5/elt-6 transcriptional
circuit and possibly many others.

Yousin Suh (Einstein College of Medicine, USA)
presented "MicroRNA, aging, and longevity". Expression levels of miRNAs in
lymphoblastoid cell lines and plasma are associated with longevity in humans
and that some of these miRNAs are known to target conserved pathways of aging,
including the IGF signaling pathway.

Alex Maslov (Einstein College of Medicine, USA)
presented "DNA damage in normal and premature aging: impact on the aging
epigenome". Quantitative long-range PCR is a reliable method for quantitative
assessment of DNA damageю Normally, aged animals demonstrate an
increased level of DNA damage across the genome.

Julia Hoffmann and Thomas Roeder (University of
Kiel, Germany) reported that mild sir2 overexpression in the
fat body of D. melanogaster extends life span and reduces
relative body fat content in both males and females. The response to dietary
restriction (DR) is gender-specific: DR reduces relative body fat content in
females but increases it in males. Regulated gene sets of males and females
show a very small overlap.

CONFERENCE PICTURES

Alexey Moskalev (co-chair of the Organizing committee), Mikhail A. Batin (co-chair of the
Organizing committee) and Robert Shmookler Reis